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Os resultados encontrados nessa dissertação permitem as seguintes conclusões:
1. Identificamos cinco kits comerciais como principais opções disponíveis para monitoramento de BCR-ABL1 na LMC. Avaliamos individualmente cada um deles, com relação as suas características analíticas e especificações técnicas. Abordamos ainda, vantagens, desvantagens e informações técnicas inerentes, que possibilitam a tomada de decisão na escolha de um teste para rotina laboratorial;
2. A partir da comparação da avaliação técnica e operacional entre os ensaios GeneXpert® BCR-ABL eBCR-ABL1 Quant RUO™ com a metodologia referência proposta por Susan Branford e Timothy Hughes para o monitoramento de DRM em pacientes com LMC, observamos que as metodologias apresentaram resultados adequados, com o ensaio BCR-ABL1 Quant RUO™ mostrando-se mais adequado para monitoramento de pacientes que atingiram RMM mais profundas;
3. Para alinhar os resultado à IS, determinamos um FC específico para o laboratório, através de uma das estratégias recomendadas para métodos de quantificação de BCR-ABL1, utilizando painéis de referência secundários disponíveis comercialmente. Ainda, confirmamos a estabilidade deste FC a fim de padronizar os resultados com a IS;
4. Determinamos as isoformas do transcrito BCR-ABL1 através da técnica de Eletroforese Capilar (EC) a partir do amplicon do kit da Asuragen identificando as isoformas mais prevalentes em pacientes com LMC. A partir disso, realizamos a comparação dos resultados concordantes e discordantes da metodologia de referência, não evidenciando um perfil característico entre os ensaios testados;
5. Diante dos aspectos técnicos e operacionais avaliados, observamos que o ensaio GeneXpert® BCR-ABL pode ser considerado um bom teste primário a ser
realizado em pacientes que iniciaram o tratamento recentemente ou em casos onde há suspeita da perda de RM devido a sintomas clínicos. Já o ensaio BCR-ABL1
Quant RUO™, por ter demonstrado melhor comparabilidade com RMM em análises
de respostas moleculares profundas, poderia ser um teste secundário, a fim de confirmar resultados abaixo de uma RMM ou resultados não detectados.
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Diante dos resultados encontrados e visando o aprofundamento e melhor entendimento das relações estabelecidas para os ensaios avaliados, algumas perspectivas podem ser estabelecidas:
1. De maneira geral, aumentar o número amostral para as diferentes faixas de resposta molecular, a fim de avaliar de forma mais ampla o desempenho das metodologias. Uma das limitações impostas na análise de nossos resultados foi a não homogeneidade em todas as faixas de RMM, mesmo com uma coleta de amostras realizadas prospectivamente por quase dois anos;
2. A partir do desenvolvimento de novas metodologias mais sensíveis, como a PCR digital, avaliar novas amostras sem comparação com a metodologia referência, porém sempre com a mesma ótica apresentada neste trabalho, considerando uma relação de aplicabilidade para serviços locais;
3. Realizar o acompanhamento do paciente em relação ao ITQ utilizado, tempo de tratamento, alcance de RMM, monitoramento de DRM a partir da implantação do kit de escolha no serviço. A partir disso, observar junto aos clínicos sua funcionalidade, desempenho e ganho de qualidade na tomada de decisão.
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139 10.1 Anexo I
Parecer Consubstanciado do Comitê de Ética - Hospital de Clínicas de Porto Alegre – HCPA/ UFRGS – Projeto 1.
145 10.2 Anexo II
Parecer Consubstanciado do Comitê de Ética - Hospital de Clínicas de Porto Alegre – HCPA/ UFRGS – Projeto 2.